Heat-Shock Proteins Gene Expression in Peripheral Blood Mononuclear Cells as an Indicator of Heat Stress in Beef Calves

Abstract

This study was conducted to investigate the effect of HS on HSPs gene expression in bovine PBMCs of beef calves in in vitro and in vivo models. In the in vitro experiment, blood samples were collected from the jugular vein of five beef calves (age: 174.2 ± 5.20 days, BW: 145.2 ± 5.21 kg). In the in vivo experiment, sixteen Korean native male beef calves (age: 169.6 ± 4.60 days, BW: 136.9 ± 6.23 kg) were exposed to ambient temperature for seven days (22 to 24 °C, relative humidity 60%; temperature-humidity index (THI) = 68 to 70) and subsequently to the temperature and humidity corresponding to the target THI level for 21 days (HS). For PBMC isolation, blood samples were collected every three days. In the in vitro model, the cell viability was significantly decreased in HS groups compared with the control group (p = 0.015). The expression of HSP70 (p = 0.022), HSP90 (p = 0.003) and HSPB1 (p = 0.026) genes was increased in the HS group in in vitro model. In the in vivo experiment, the HSP70 gene expression was increased after sudden exposure to HS conditions (severe THI levels; THI = 88 to 90), whereas HSP90 and HSPB1 showed no differences among the THI groups (p > 0.05). However, in the severe THI group, the HSP70 gene expression returned to normal range after six days of continuous HS. In conclusion, the HSP70 gene plays a pivotal role in protecting cells from damage and is sensitive to HS in immune cells compared with other HSP genes in in vitro and in vivo models. In addition, the in vivo models suggest that calves exhibit active physiological mechanisms of adaptation to HS after six days of continuous exposure by regulating the HSP70 gene expression.

Keywords: PBMCs; beef calf; heat stress; heat-shock proteins.

Figure 1

Effects of heat stress on peripheral blood mononuclear cells (PBMCs) proliferation. Cell viability was measured by Cell Counting Kit-8 (CCK-8) assay. Data are presented as the means ± standard errors (n = 5 for each of three experiments). * Means with different superscripts differ significantly (p < 0.05) between two groups based on Student’s t-test.

Figure 2

mRNA expression of (a), HSP70, (b) HSP90, and (c) HSPB1 in calf PBMC cultures in an in vitro model. Data are presented as the means ± standard errors (n = 5 for each of three experiments). * Means with different superscripts differ significantly (p < 0.05) between two groups based on Student’s t-test.

Figure 3

mRNA expression of (a), HSP70, (b) HSP90, and (c) HSPB1 in the PBMCs of calves during the climate chamber experiment. Data are presented as the means ± standards error (n = 4). a, b Means with different superscripts differ significantly (p < 0.05) among the sampling dates based on Tukey’s test.

Figure 3

mRNA expression of (a), HSP70, (b) HSP90, and (c) HSPB1 in the PBMCs of calves during the climate chamber experiment. Data are presented as the means ± standards error (n = 4). a, b Means with different superscripts differ significantly (p < 0.05) among the sampling dates based on Tukey’s test.